IgGZERO (EndoS) and GlycINATOR (EndoS2) rapidly hydrolyze N-glycans on the Fc domain of IgGs. GlycINATOR removes all high-mannose glycans on human IgGs which IgGZERO does not. Both enzymes remove complex glycans whereas GlycINATOR is more effective on hybrid glycans.
Afucosylation of the IgG glycans has substantial impact on the effector functions of the antibody and the antibody dependent cytotoxicity (ADCC). To study the degree of antibody afucosylation is therefore of great importance for all antibody based therapeutics.
Genovis SmartEnzymes have been used to study the degree of afucosylation on antibodies, both in cell culture medium in clone selection and to study antibody variations in blood.
FabRICATOR is used in the subunit domain mapping / middle down approach.
As FabRICATOR only has one cleavage site regardless of incubation time, the amino acid sequence in each of the antibody fragments is exactly the same. All fragments are below 30 kD which allows for mass spectrometric determination of major glycans, degree of fucosylation and many of the post translation modifications. As ionization efficiency is mostly related to the protein sequence, intensities can be considered quantitative which allows for rapid glycan content determination.
The endoglycosidase IgGZERO (EndoS) has been put forward as a tool for enhancing the therapeutic efficacy of monoclonal antibody based products. In a paper in Science Magazine by Dalziel et al. explains the potential use of IgGZERO (EndoS).